We are continuing our efforts in characterizing the lens and processes that may occur in cataract development. There are three aims in our research effort: determination of the sequences of crystallins, development of the in vitro lens incubation system to test anticataract agents, and the examination of stress on lenses and lens crystallins. We have now sequenced the human gammaS-crystallin by a combination of cDNA sequencing, electrospray ionization mass spectrometry, and fast atom bombardment mass spectrometry. The promoter region of mouse betaB2- crystallin has been identified and sequenced. The binding sites for various transcription factors have been determined. The transcription of betaB-2-crystallin appears to be under the control of the Pax 6 protein, which is essential for eye development. The lens work using the in vitro system has shown that the monkey lens has a much more stable glutathione content than the young rat lens when placed into organ culture. Experiments have indicated that the reason for this is the slower rate of growth of older lenses. Older lenses also appear to have less RNA than more rapidly growing ones. It was determined that most if not all of the protein synthesis in a monkey lens occurs in the epithelium and elongating fiber cells. Three enzymes involved in the protection of the lens during oxidative stress-catalase, glutathione peroxidase, and superoxide dismutase--have been studied with organ-cultured monkey lenses. The changes in these enzymes are dependent on the amount of oxidative stress, but the experiments suggest coordinated expression of these enzymes in the lens. The effects of environmental stress have been studied using cell culture systems. The cell line used in these studies constitutively expressed alphaB-crystallin. Effects of oxidative stress and heat shock were investigated on this crystallin, which is thought to be a molecular chaperone.